Detection and quantification of the toxic marine microalgae Karlodinium veneficum and Karlodinium armiger using recombinase polymerase amplification and enzyme-linked oligonucleotide assay
Ver/Abrir
Autor/a
Toldrà, Anna
Jauset-Rubio, Míriam
Andree, Karl B.
Katakis, Ioanis
O'Sullivan, Ciara K.
Fecha de publicación
2018-07-28ISSN
0003-2670
Resumen
Karlodinium is a dinoflagellate responsible for fish-killing events worldwide. In Alfacs Bay (NW Mediterranean Sea), the presence of two Karlodinium species (K. veneficum and K. armiger) with different toxicities has been reported. This work presents a method that combines recombinase polymerase amplification (RPA) with an enzyme-linked oligonucleotide assay (ELONA) to identify, discriminate and quantify these two species. The system was characterised using synthetic DNA and genomic DNA, and the specificity was confirmed by cross-reactivity experiments. Calibration curves were constructed using 10-fold dilutions of cultured cells, attaining a limit of detection of around 50,000 cells/L, far below the Karlodinium spp. alert threshold (200,000 cells/L). Finally, the assay was applied to spiked seawater samples, showing an excellent correlation with the spiking levels and light microscopy counts. This approach is more rapid, specific and user-friendly than traditional microscopy techniques, and shows great promise for the surveillance and management of harmful algal blooms.
Tipo de documento
Artículo
Versión del documento
Versión aceptada
Lengua
English
Materias (CDU)
63 - Agricultura. Silvicultura. Zootecnia. Caza. Pesca
Páginas
19
Publicado por
Elsevier
Publicado en
Analytica Chimica Acta
Citación
Toldrà, Anna, Míriam Jauset-Rubio, Karl B. Andree, Margarita Fernández-Tejedor, Jorge Diogène, Ioanis Katakis, Ciara K. O'Sullivan, and Mònica Campàs. 2018. "Detection And Quantification Of The Toxic Marine Microalgae Karlodinium Veneficum And Karlodinium Armiger Using Recombinase Polymerase Amplification And Enzyme-Linked Oligonucleotide Assay". Analytica Chimica Acta 1039: 140-148. Elsevier BV. doi:10.1016/j.aca.2018.07.057.
Número del acuerdo de la subvención
MINECO/Programa Estatal de I+D+I orientada a los retos de la sociedad/BIO2017-87946-C2-2-R/ES/DISPOSITIVOS BIOANALITICOS PARA LA DETECCION RAPIDA Y RENTABLE DE GAMBIERDISCUS Y CIGUATOXINAS/CIGUASENSING
MINECO/Programa Estatal de I+D+I orientada a los retos de la sociedad/BIO2014-56024-C2-2-R/ES/MICROSISTEMAS PARA LA DETECCION RAPIDA, FIABLE Y RENTABLE DE MICROALGAS TOXICAS IN SITU Y A TIEMPO REAL/SEASENSING
Program
Aigües Marines i Continentals
Aqüicultura
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